Bacterial Adaptive Memory in Methicillin-Resistant Staphylococcus aureus from Endotracheal Tubes.

OBJECTIVES: To evaluate the expression dynamics of biofilm genes in methicillin-resistant Staphylococcus aureus (MRSA) retrieved from endotracheal tubes (ETT) and to determine how gene regulation is attenuated in vitro where host-environmental factors are no longer present. METHODS: Biofilm was grown (24 h) in tryptic broth soy plus 0.25% glucose for a clinical MRSA isolate in planktonic state and after sessile growth named ETT-MRSA (S2, S3, S4, S5, S6, S7). Gene expression of five biofilm-related genes (icaC, clfB, ebps, fnbB, and RNA III) was assessed consecutively from day 1 to day 4 after ETT growth through real-time PCR. 16S rRNA was used as a control. RESULTS: The MRSA isolates retrieved from ETT were capable of producing biofilms dependent on ica. The gene expression dynamics of ETT-MRSA changed progressively compared to planktonic MRSA gene expression under both ambient air (p < 0.001) and ambient air with 5% CO2 (p < 0.001). Dynamic assessment of icaC expression in both atmospheric conditions showed progressive downregulation in vitro compared to in vivo ETT biofilms. The expression patterns of clfB and ebps genes were similar to icaC. In contrast, the expression of the RNA III gene showed progressive upregulation from day 1 to day 4 (p < 0.001). CONCLUSIONS: MRSA loses its biofilm gene expression in vitro, by adaptive features across multiple generations, as evidenced by the progressive downregulation of icaC and upregulation of RNA III. These findings underscore the significance of host-environment dependence in regulating bacterial biofilm genes, highlighting its importance in diagnostics. Bacterial strains lose their host-specific characteristics as they are cultured in vitro.

A long-lasting porcine model of ARDS caused by pneumonia and ventilator-induced lung injury.

BACKGROUND: Animal models of acute respiratory distress syndrome (ARDS) do not completely resemble human ARDS, struggling translational research. We aimed to characterize a porcine model of ARDS induced by pneumonia-the most common risk factor in humans-and analyze the additional effect of ventilator-induced lung injury (VILI). METHODS: Bronchoscopy-guided instillation of a multidrug-resistant Pseudomonas aeruginosa strain was performed in ten healthy pigs. In six animals (pneumonia-with-VILI group), pulmonary damage was further increased by VILI applied 3 h before instillation and until ARDS was diagnosed by PaO2/FiO2 < 150 mmHg. Four animals (pneumonia-without-VILI group) were protectively ventilated 3 h before inoculum and thereafter. Gas exchange, respiratory mechanics, hemodynamics, microbiological studies and inflammatory markers were analyzed during the 96-h experiment. During necropsy, lobar samples were also analyzed. RESULTS: All animals from pneumonia-with-VILI group reached Berlin criteria for ARDS diagnosis until the end of experiment. The mean duration under ARDS diagnosis was 46.8 ± 7.7 h; the lowest PaO2/FiO2 was 83 ± 5.45 mmHg. The group of pigs that were not subjected to VILI did not meet ARDS criteria, even when presenting with bilateral pneumonia. Animals developing ARDS presented hemodynamic instability as well as severe hypercapnia despite high-minute ventilation. Unlike the pneumonia-without-VILI group, the ARDS animals presented lower static compliance (p = 0.011) and increased pulmonary permeability (p = 0.013). The highest burden of P. aeruginosa was found at pneumonia diagnosis in all animals, as well as a high inflammatory response shown by a release of interleukin (IL)-6 and IL-8. At histological examination, only animals comprising the pneumonia-with-VILI group presented signs consistent with diffuse alveolar damage. CONCLUSIONS: In conclusion, we established an accurate pulmonary sepsis-induced ARDS model.

The value of biofilm testing to guide antimicrobial stewardship in chronic respiratory diseases.

Introduction: Biofilm production is an important yet currently overlooked aspect of diagnostic microbiology that has implications for antimicrobial stewardship. In this study, we aimed to validate and identify additional applications of the BioFilm Ring Test® (BRT) for Pseudomonas aeruginosa (PA) isolates from patients with bronchiectasis (BE). Materials and methods: Sputa were collected from BE patients who had at least one PA positive culture in the previous year. We processed the sputa to isolate both mucoid and non-mucoid PA, and determined their susceptibility pattern, mucA gene status, and presence of ciprofloxacin mutations in QRDR genes. The Biofilm production index (BPI) was obtained at 5 and 24 hours. Biofilms were imaged using Gram staining. Results: We collected 69 PA isolates, including 33 mucoid and 36 non-mucoid. A BPI value below 14.75 at 5 hours predicted the mucoid PA phenotype with 64% sensitivity and 72% specificity. Conclusion: Overall, our findings suggest that the fitness-cost associated with the mucoid phenotype or ciprofloxacin resistance is shown through a time-dependent BPI profile. The BRT has the potential to reveal biofilm features with clinical implications.

Inhaled amikacin for pneumonia treatment and dissemination prevention: an experimental model of severe monolateral Pseudomonas aeruginosa pneumonia.

BACKGROUND: Pseudomonas aeruginosa pneumonia is commonly treated with systemic antibiotics to ensure adequate treatment of multidrug resistant (MDR) bacteria. However, intravenous (IV) antibiotics often achieve suboptimal pulmonary concentrations. We therefore aimed to evaluate the effect of inhaled amikacin (AMK) plus IV meropenem (MEM) on bactericidal efficacy in a swine model of monolateral MDR P. aeruginosa pneumonia. METHODS: We ventilated 18 pigs with monolateral MDR P. aeruginosa pneumonia for up to 102 h. At 24 h after the bacterial challenge, the animals were randomized to receive 72 h of treatment with either inhaled saline (control), IV MEM only, or IV-MEM plus inhaled AMK (MEM + AMK). We dosed IV MEM at 25 mg/kg every 8 h and inhaled AMK at 400 mg every 12 h. The primary outcomes were the P. aeruginosa burden and histopathological injury in lung tissue. Secondary outcomes included the P. aeruginosa burden in tracheal secretions and bronchoalveolar lavage fluid, the development of antibiotic resistance, the antibiotic distribution, and the levels of inflammatory markers. RESULTS: The median (25-75th percentile) P. aeruginosa lung burden for animals in the control, MEM only, and MEM + AMK groups was 2.91 (1.75-5.69), 0.72 (0.12-3.35), and 0.90 (0-4.55) log10 CFU/g (p = 0.009). Inhaled therapy had no effect on preventing dissemination compared to systemic monotherapy, but it did have significantly higher bactericidal efficacy in tracheal secretions only. Remarkably, the minimum inhibitory concentration of MEM increased to > 32 mg/L after 72-h exposure to monotherapy in 83% of animals, while the addition of AMK prevented this increase (p = 0.037). Adjunctive therapy also slightly affected interleukin-1ß downregulation. Despite finding high AMK concentrations in pulmonary samples, we found no paired differences in the epithelial lining fluid concentration between infected and non-infected lungs. Finally, a non-significant trend was observed for higher amikacin penetration in low-affected lung areas. CONCLUSIONS: In a swine model of monolateral MDR P. aeruginosa pneumonia, resistant to the inhaled AMK and susceptible to the IV antibiotic, the use of AMK as an adjuvant treatment offered no benefits for either the colonization of pulmonary tissue or the prevention of pathogen dissemination. However, inhaled AMK improved bacterial eradication in the proximal airways and hindered antibiotic resistance.

Cirugía plástica periodontal. Aplicación de la técnica de colgajo semilunar / Periodontal plastic surgery. Application of the semilunar flap technique / Cirurgia plástica periodontal. Aplicação da técnica de retalho semilunar

RESUMEN Introducción: cada día es más frecuente observar la preocupación de los pacientes por la presencia de trastornos mucogingivales, y dentro de estos, uno de los más comunes es la recesión periodontal. Esta puede acompañarse de sensibilidad radicular, mayor prevalencia de caries y abrasiones cervicales. Presentación de caso: el caso que presentamos se trató de un paciente de 25 años de edad del sexo masculino que acudió a nuestra consulta preocupado por su estética y sensibilidad dental a los cambios térmicos. El mismo presentaba una recesión periodontal de 3 mm a nivel del 13. Discusión: decidimos darle solución al problema con la técnica de colgajo semilunar, cumpliéndose el objetivo del trabajo al quedar resueltos los problemas de estéticas y sensibilidad del paciente una vez cubierta la raíz y aumentado el ancho la encía insertada. Conclusiones: al examen clínico los resultados mostraron un recubrimiento radicular, aumento del grosor de encía insertada, disminución de sensibilidad dental y estética aceptada por el paciente.

ABSTRACT Introduction: every day it is more frequent to observe the concern of patients for the presence of mucogingival disorders, and within these, one of the most common is periodontal recession. This can be accompanied by root sensitivity, increased prevalence of caries and cervical abrasions. Case presentation: the case we presented was a 25-year-old male patient who came to our office concerned about his aesthetics and dental sensitivity to thermal changes. It presented a periodontal recession of 3 mm at the level of 13. Discussion: we decided to solve the problem with the semilunar flap technique, fulfilling the objective of the work by solving the problems of aesthetics and sensitivity of the patient once the root is covered and the width of the inserted gum is increased. Conclusions: on clinical examination the results showed a root coating, increase in the thickness of the inserted gum, decrease in dental sensitivity and aesthetics accepted by the patient.

RESUMO Introdução: a cada dia é mais frequente observar a preocupação dos pacientes com a presença de distúrbios mucogingival, e dentro destes, um dos mais comuns é a recessão periodontal. Isso pode ser acompanhado de sensibilidade radicular, aumento da prevalência de cárter e abrasões cervicais. Apresentação do caso: o caso que apresentamos foi um paciente do sexo masculino de 25 anos que veio ao nosso consultório preocupado com sua estética e sensibilidade dentária às mudanças térmicas. Apresentou uma recessão periodontal de 3 mm ao nível de 13. Discussão: decidimos resolver o problema com a técnica de retalho semilunar, cumprindo o objetivo do trabalho resolvendo os problemas da estética e sensibilidade do paciente uma vez que a raiz é coberta e a largura da gengiva inserida é aumentada. Conclusões: no exame clínico os resultados mostraram revestimento radicular, aumento da espessura da gengiva inserida, diminuição da sensibilidade dentária e estética aceita pelo paciente.

Resistance mechanisms and molecular epidemiology of Pseudomonas aeruginosa strains from patients with bronchiectasis.

BACKGROUND: Non-cystic fibrosis bronchiectasis (BE) is a chronic structural lung condition that facilitates chronic colonization by different microorganisms and courses with recurrent respiratory infections and frequent exacerbations. One of the main pathogens involved in BE is Pseudomonas aeruginosa. OBJECTIVES: To determine the molecular mechanisms of resistance and the molecular epidemiology of P. aeruginosa strains isolated from patients with BE. METHODS: A total of 43 strains of P. aeruginosa were isolated from the sputum of BE patients. Susceptibility to the following antimicrobials was analysed: ciprofloxacin, meropenem, imipenem, amikacin, tobramycin, aztreonam, piperacillin/tazobactam, ceftazidime, ceftazidime/avibactam, ceftolozane/tazobactam, cefepime and colistin. The resistance mechanisms present in each strain were assessed by PCR, sequencing and quantitative RT-PCR. Molecular epidemiology was determined by MLST. Phylogenetic analysis was carried out using the eBURST algorithm. RESULTS: High levels of resistance to ciprofloxacin (44.19%) were found. Mutations in the gyrA, gyrB, parC and parE genes were detected in ciprofloxacin-resistant P. aeruginosa strains. The number of mutated QRDR genes was related to increased MIC. Different ß-lactamases were detected: blaOXA50, blaGES-2, blaIMI-2 and blaGIM-1. The aac(3)-Ia, aac(3)-Ic, aac(6″)-Ib and ant(2″)-Ia genes were associated with aminoglycoside-resistant strains. The gene expression analysis showed overproduction of the MexAB-OprM efflux system (46.5%) over the other efflux system. The most frequently detected clones were ST619, ST676, ST532 and ST109. CONCLUSIONS: Resistance to first-line antimicrobials recommended in BE guidelines could threaten the treatment of BE and the eradication of P. aeruginosa, contributing to chronic infection.

Cirugía plástica periodontal. Aplicación de la técnica de colgajo semilunar / Periodontal plastic surgery. Application of the semilunar flap technique

Introducción: cada día es más frecuente observar la preocupación de los pacientes por la presencia de trastornos mucogingivales, y dentro de estos, uno de los más comunes es la recesión periodontal. Esta puede acompañarse de sensibilidad radicular, mayor prevalencia de caries y abrasiones cervicales. Presentación de caso: el caso que presentamos se trató de un paciente de 25 años de edad del sexo masculino que acudió a nuestra consulta preocupado por su estética y sensibilidad dental a los cambios térmicos. El mismo presentaba una recesión periodontal de 3 mm a nivel del 13. Discusión: decidimos darle solución al problema con la técnica de colgajo semilunar, cumpliéndose el objetivo del trabajo al quedar resueltos los problemas de estéticas y sensibilidad del paciente una vez cubierta la raíz y aumentado el ancho la encía insertada. Conclusiones: al examen clínico los resultados mostraron un recubrimiento radicular, aumento del grosor de encía insertada, disminución de sensibilidad dental y estética aceptada por el paciente(AU)

Introduction: every day it is more frequent to observe the concern of patients for the presence of mucogingival disorders, and within these, one of the most common is periodontal recession. This can be accompanied by root sensitivity, increased prevalence of caries and cervical abrasions. Case presentation: the case we presented was a 25-year-old male patient who came to our office concerned about his aesthetics and dental sensitivity to thermal changes. Itpresented a periodontal recession of 3 mm at the level of 13. Discussion: we decided to solve the problem with the semilunar flap technique, fulfillingthe objective of the work by solving the problems of aesthetics and sensitivity of the patient once the root is covered and the width of the inserted gum is increased. Conclusions: on clinical examination the results showed a root coating, increase in the thickness of the inserted gum, decrease in dental sensitivity and aesthetics accepted by the patient(AU)

Aplicación del plasma rico en plaquetas en la enfermedad periodontal / Application of platelet-rich plasma in periodontal disease / Aplicação de plasma rico em plaquetas na doença periodontal

RESUMEN La periodontitis es una afección crónica que afecta los tejidos de soporte del diente lo que conlleva en ocasiones a la pérdida del mismo. El objetivo fue evaluar la eficacia y seguridad del plasma rico en plaquetas en el tratamiento de la periodontitis crónica. Se realizó un ensayo clínico en fase II b, controlado, aleatorizado, con evaluación a ciegas por terceros. La muestra abarcó 70 pacientes portadores de periodontitis crónica que acudieron a consulta de Periodoncia de la Clínica Universitaria de Especialidades Estomatológicas "Manuel Cedeño" entre enero 2017 a diciembre del 2019. Los individuos quedaron distribuidos aleatoriamente en: grupo A o experimental tratado con plasma rico en plaquetas y grupo B o control con tratamiento convencional. Las variables fueron: edad, sexo, inflamación de la encía, profundidad de las bolsas periodontales, pérdida de inserción periodontal, tiempo de evolución, efecto terapéutico y eventos adversos. Los resultados mostraron que a los 12 meses del tratamiento curaron 34 pacientes del grupo experimental (97,14 %) y 33 pacientes del grupo control (94,39 %). No se detectaron eventos adversos. Se llegó a la conclusión que el tratamiento con plasma rico en plaquetas en la periodontitis crónica demostró eficacia terapéutica y seguridad sin diferencias significativas con respecto al tratamiento convencional.

ABSTRACT Periodontitis is a chronic condition that affects the supporting tissues of the tooth, which sometimes leads to tooth loss. The objective was to evaluate the efficacy and safety of platelet-rich plasma in the treatment of chronic periodontitis. A randomized, controlled, phase II b clinical trial was conducted with blind third-party evaluation. The sample comprised 70 patients with chronic periodontitis who attended the Periodontics consultation of the "Manuel Cedeño" University Clinic of Stomatological Specialties between January 2017 and December 2019. The individuals were randomly distributed into: group A or experimental group treated with plasma rich in platelets and group B or control with conventional treatment. The variables were: age, sex, inflammation of the gingiva, depth of periodontal pockets, loss of periodontal attachment, time of evolution, therapeutic effect and adverse events. The results showed that at 12 months of treatment 34 patients in the experimental group (97,14 %) and 33 patients in the control group (94,39 %) cured. No adverse events were detected. It was concluded that treatment with platelet-rich plasma in chronic periodontitis demonstrated therapeutic efficacy and safety without significant differences with respect to conventional treatment.

RESUMO A periodontite é uma condição crônica que afeta os tecidos de suporte do dente, o que às vezes leva à perda do dente. O objetivo foi avaliar a eficácia e segurança do plasma rico em plaquetas no tratamento da periodontite crônica. Um ensaio clínico randomizado, controlado, fase II b foi conduzido com avaliação cega por terceiros. A amostra foi composta por 70 pacientes com periodontite crônica que compareceram à consulta de Periodontia da Clínica Universitária de Especialidades Estomatológicas "Manuel Cedeño" entre janeiro de 2017 e dezembro de 2019. Os indivíduos foram distribuídos aleatoriamente em: grupo A ou grupo experimental tratado com plasma rico em plaquetas e grupo B ou controle com tratamento convencional. As variáveis ​​foram: idade, sexo, inflamação da gengiva, profundidade das bolsas periodontais, perda da inserção periodontal, tempo de evolução, efeito terapêutico e eventos adversos. Os resultados mostraram que, aos 12 meses de tratamento, 34 pacientes do grupo experimental (97,14%) e 33 pacientes do grupo controle (94,39%) curaram. Nenhum evento adverso foi detectado. Concluiu-se que o tratamento com plasma rico em plaquetas na periodontite crônica demonstrou eficácia terapêutica e segurança sem diferenças significativas em relação ao tratamento convencional.

Daidzein modulates cocaine-reinforcing effects and cue-induced cocaine reinstatement in CD-1 male mice.

RATIONALE: Cocaine addiction is a chronic relapsing disorder that lacks of an effective treatment. Isoflavones are a family of compounds present in different plants and vegetables like soybeans that share a common chemical structure. Previous studies have described that synthetic derivatives from the natural isoflavone daidzin can modulate cocaine addiction, by a mechanism suggested to involve aldehyde-dehydrogenase (ALDH) activities. OBJECTIVES: Based on these previous studies, we investigated the effects of three natural isoflavones, daidzin, daidzein, and genistein, on the modulation of the cocaine reinforcing effects and on cue-induced reinstatement in an operant mouse model of cocaine self-administration. RESULTS: Chronic treatment with daidzein or genistein decreased operant responding to obtain cocaine intravenous infusions. On the other hand, daidzein and daidzin, but not genistein, were effective in decreasing cue-induced cocaine reinstatement. Complementary studies revealed that daidzein effects on cocaine reinforcement were mediated through a mechanism that involved dopamine type-2/3 receptors (DA-D2/3) activities. CONCLUSIONS: Our results suggest that these natural compounds alone or in combination can be a potential therapeutic approach for cocaine addiction. Further clinical studies are required in order to ascertain their potential therapeutic use.

Short-Term Effects of Appropriate Empirical Antimicrobial Treatment with Ceftolozane/Tazobactam in a Swine Model of Nosocomial Pneumonia.

The rising frequency of multidrug-resistant and extensively drug-resistant (MDR/XDR) pathogens is making more frequent the inappropriate empirical antimicrobial therapy (IEAT) in nosocomial pneumonia, which is associated with increased mortality. We aim to determine the short-term benefits of appropriate empirical antimicrobial treatment (AEAT) with ceftolozane/tazobactam (C/T) compared with IEAT with piperacillin/tazobactam (TZP) in MDR Pseudomonas aeruginosa pneumonia. Twenty-one pigs with pneumonia caused by an XDR P. aeruginosa strain (susceptible to C/T but resistant to TZP) were ventilated for up to 72 h. Twenty-four hours after bacterial challenge, animals were randomized to receive 2-day treatment with either intravenous saline (untreated) or 25 to 50 mg of C/T per kg body weight (AEAT) or 200 to 225 mg of TZP per kg (IEAT) every 8 h. The primary outcome was the P. aeruginosa burden in lung tissue and the histopathology injury. P. aeruginosa burden in tracheal secretions and bronchoalveolar lavage (BAL) fluid, the development of antibiotic resistance, and inflammatory markers were secondary outcomes. Overall, P. aeruginosa lung burden was 5.30 (range, 4.00 to 6.30), 4.04 (3.64 to 4.51), and 4.04 (3.05 to 4.88) log10CFU/g in the untreated, AEAT, and IEAT groups, respectively (P = 0.299), without histopathological differences (P = 0.556). In contrast, in tracheal secretions (P < 0.001) and BAL fluid (P = 0.002), bactericidal efficacy was higher in the AEAT group. An increased MIC to TZP was found in 3 animals, while resistance to C/T did not develop. Interleukin-1ß (IL-1ß) was significantly downregulated by AEAT in comparison to other groups (P = 0.031). In a mechanically ventilated swine model of XDR P. aeruginosa pneumonia, appropriate initial treatment with C/T decreased respiratory secretions' bacterial burden, prevented development of resistance, achieved the pharmacodynamic target, and may have reduced systemic inflammation. However, after only 2 days of treatment, P. aeruginosa tissue concentrations were moderately affected.

Aplicación del plasma rico en plaquetas en la enfermedad periodontal / Application of platelet-rich plasma in periodontal disease

La periodontitis es una afección crónica que afecta los tejidos de soporte del diente lo que conlleva en ocasiones a la pérdida del mismo. El objetivo fue evaluar la eficacia y seguridad del plasma rico en plaquetas en el tratamiento de la periodontitis crónica. Se realizó un ensayo clínico en fase II b, controlado, aleatorizado, con evaluación a ciegas por terceros. La muestra abarcó 70 pacientes portadores de periodontitis crónica que acudieron a consulta de Periodoncia de la Clínica Universitaria de Especialidades Estomatológicas "Manuel Cedeño" entre enero 2017 a diciembre del 2019. Los individuos quedaron distribuidos aleatoriamente en: grupo A o experimental tratado con plasma rico en plaquetas y grupo B o control con tratamiento convencional. Las variables fueron: edad, sexo, inflamación de la encía, profundidad de las bolsas periodontales, pérdida de inserción periodontal, tiempo de evolución, efecto terapéutico y eventos adversos. Los resultados mostraron que a los 12 meses del tratamiento curaron 34 pacientes del grupo experimental (97,14 por ciento) y 33 pacientes del grupo control (94,39 por ciento). No se detectaron eventos adversos. Se llegó a la conclusión que el tratamiento con plasma rico en plaquetas en la periodontitis crónica demostró eficacia terapéutica y seguridad sin diferencias significativas con respecto al tratamiento convencional(AU)

Periodontitis is a chronic condition that affects the supporting tissues of the tooth, which sometimes leads to tooth loss. The objective was to evaluate the efficacy and safety of platelet-rich plasma in the treatment of chronic periodontitis. A randomized, controlled, phase II b clinical trial was conducted with blind third-party evaluation. The sample comprised 70 patients with chronic periodontitis who attended the Periodontics consultation of the "Manuel Cedeño" University Clinic of Stomatological Specialties between January 2017 and December 2019. The individuals were randomly distributed into: group A or experimental group treated with plasma rich in platelets and group B or control with conventional treatment. The variables were: age, sex, inflammation of the gingiva, depth of periodontal pockets, loss of periodontal attachment, time of evolution, therapeutic effect and adverse events. The results showed that at 12 months of treatment 34 patients in the experimental group (97,14 percent) and 33 patients in the control group (94,39 percent) cured. No adverse events were detected. It was concluded that treatment with platelet-rich plasma in chronic periodontitis demonstrated therapeutic efficacy and safety without significant differences with respect to conventional treatment(EU)

Molecular characterization of methicillin-resistant Staphylococcus aureus clinical strains from the endotracheal tubes of patients with nosocomial pneumonia.

BACKGROUND: Among all cases of nosocomial pneumonia, Staphylococcus aureus is the second most prevalent pathogen (17.8%). In Europe, 29.9% of the isolates are oxacillin-resistant. The changing epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) nosocomial infections and the decreasing susceptibility to first-line antibiotics leave clinicians with few therapeutic options. The objective of our study was to determine the antimicrobial susceptibility, the associated molecular mechanisms of resistance and the epidemiological relatedness of MRSA strains isolated from the endotracheal tubes (ETT) of intubated critically ill patients in the intensive care unit (ICU) with nosocomial pneumonia caused by Staphylococcus aureus. METHODS: The antimicrobial susceptibility to vancomycin, linezolid, ciprofloxacin, clindamycin, erythromycin, chloramphenicol, fusidic acid, gentamicin, quinupristin-dalfopristin, rifampicin, sulfamethoxazole/trimethoprim, and tetracycline were measured. Resistance mechanisms were then analyzed by polymerase chain reaction and sequencing. Molecular epidemiology was carried out by multi-locus sequence typing. RESULTS: S. aureus isolates were resistant to ciprofloxacin, erythromycin, gentamicin, tetracycline, clindamycin, and fusidic acid. The most frequent mutations in quinolone-resistant S. aureus strains were S84L in the gyrA gene, V511A in the gyrB gene, S144P in the grlA gene, and K401R/E in the grlB gene. Strains resistant to erythromycin carried the ermC, ermA, and msrA genes; the same ermC and ermA genes were detected in strains resistant to clindamycin. The aac(6')-aph(2″) gene was related to gentamicin resistance, while resistance to tetracycline was related to tetK (efflux pump). The fusB gene was detected in the strain resistant to fusidic acid. The most frequent sequence types were ST22, ST8, and ST217, which were distributed in four clonal complexes (CC5, CC22, CC45, and CC59). CONCLUSIONS: High levels of resistance to second-line antimicrobials threatens the treatment of nosocomial respiratory infections due to methicillin-resistant S. aureus with decreased susceptibility to linezolid and vancomycin. The wide genotypic diversity found reinforces the central role of ICU infection control in preventing nosocomial transmission.

Prevención de enfermedades periodontales. Métodos mecánicos de control de placa dentobacteriana

Las enfermedades periodontales afectan a las encías y las estructuras que soportan los dientes. Hablamos de la gingivitis y la periodontitis. Algunos de sus síntomas son la inflamación y el sangrado de las encías. La causa principal de desarrollo de estas afecciones suele ser la falta de higiene, que favorece la acumulación de placa dentobacteriana entre los dientes y las encías. Con el objetivo de describir los diferentes métodos que existen para remover la placa dentobacteriana causa principal de las enfermedades periodontales, se revisaron 6 bibliografías que incluyen documentos en bibliotecas, y portales digitales en internet, haciéndose uso de la información más actualizada y relacionada con el tema. Se llegó a la conclusión de que con la descripción de los métodos de control de la placa dentobacteriana estaremos capacitados para prevenir las enfermedades periodontales. .

Periodontal diseases affect the gums and the structures that support the teeth. We are talking about gingivitis and periodontitis. Some of its symptoms are inflammation and bleeding of the gums. The main cause of development of these conditions is usually the lack of hygiene, which favors the accumulation of dentobacterial plaque between the teeth and the gums. In order to describe the different methods that exist to remove the plaque dentobacteriana main cause of periodontal diseases, we reviewed 6 bibliographies that include documents in libraries, and digital portals on the Internet, making use of the most updated information related to the subject. It was concluded that with the description of the methods of control of the plaque dentobacteriana we will be trained to prevent periodontal diseases.

Prevención de enfermedades periodontales. Métodos mecánicos de control de placa dentobacteriana / Prevention of periodontal diseases. Mechanical methods of dentobacterial plaque control

Las enfermedades periodontales afectan a las encías y las estructuras que soportan los dientes. Hablamos de la gingivitis y la periodontitis. Algunos de sus síntomas son la inflamación y el sangrado de las encías. La causa principal de desarrollo de estas afecciones suele ser la falta de higiene, que favorece la acumulación de placa dentobacteriana entre los dientes y las encías. Con el objetivo de describir los diferentes métodos que existen para remover la placa dentobacteriana causa principal de las enfermedades periodontales, se revisaron 6 bibliografías que incluyen documentos en bibliotecas, y portales digitales en internet, haciéndose uso de la información más actualizada y relacionada con el tema. Se llegó a la conclusión de que con la descripción de los métodos de control de la placa dentobacteriana estaremos capacitados para prevenir las enfermedades periodontales(AU)

Periodontal diseases affect the gums and the structures that support the teeth. We are talking about gingivitis and periodontitis. Some of its symptoms are inflammation and bleeding of the gums. The main cause of development of these conditions is usually the lack of hygiene, which favors the accumulation of dentobacterial plaque between the teeth and the gums. In order to describe the different methods that exist to remove the plaque dentobacteriana main cause of periodontal diseases, we reviewed 6 bibliographies that include documents in libraries, and digital portals on the Internet, making use of the most updated information related to the subject. It was concluded that with the description of the methods of control of the plaque dentobacteriana we will be trained to prevent periodontal diseases(EU)

Melanin-Concentrating Hormone acts through hypothalamic kappa opioid system and p70S6K to stimulate acute food intake.

Melanin-Concentrating Hormone (MCH) is one of the most relevant orexigenic factors specifically located in the lateral hypothalamic area (LHA), with its physiological relevance demonstrated in studies using several genetically manipulated mice models. However, the central mechanisms controlling MCH-induced hyperphagia remain largely uncharacterized. Here, we show that central injection of MCH in mice deficient for kappa opoid receptor (k-OR) failed to stimulate feeding. To determine the hypothalamic area responsible for this MCH/k-OR interaction, we performed virogenetic studies and found that downregulation of k-OR by adeno-associated viruses (shOprk1-AAV) in LHA, but not in other hypothalamic nuclei, was sufficient to block MCH-induced food intake. Next, we sought to investigate the molecular signaling pathway within the LHA that mediates acute central MCH stimulation of food intake. We found that MCH activates k-OR and that increased levels of phosphorylated extracellular signal regulated kinase (ERK) are associated with downregulation of phospho-S6 Ribosomal Protein. This effect was prevented when a pharmacological inhibitor of k-OR was co-administered with MCH. Finally, the specific activation of the direct upstream regulator of S6 (p70S6K) in the LHA attenuated MCH-stimulated food consumption. Our results reveal that lateral hypothalamic k-OR system modulates the orexigenic action of MCH via the p70S6K/S6 pathway.

The endocannabinoid hydrolysis inhibitor SA-57: Intrinsic antinociceptive effects, augmented morphine-induced antinociception, and attenuated heroin seeking behavior in mice.

Although opioids are highly efficacious analgesics, their abuse potential and other untoward side effects diminish their therapeutic utility. The addition of non-opioid analgesics offers a promising strategy to reduce required antinociceptive opioid doses that concomitantly reduce opioid-related side effects. Inhibitors of the primary endocannabinoid catabolic enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) show opioid-sparing effects in preclinical models of pain. As simultaneous inhibition of these enzymes elicits enhanced antinociceptive effects compared with single enzyme inhibition, the present study tested whether the dual FAAH-MAGL inhibitor SA-57 [4-[2-(4-chlorophenyl)ethyl]-1-piperidinecarboxylic acid 2-(methylamino)-2-oxoethyl ester] produces morphine-sparing antinociceptive effects, without major side effects associated with either drug class. SA-57 dose-dependently reversed mechanical allodynia in the constriction injury (CCI) of the sciatic nerve model of neuropathic pain and carrageenan inflammatory pain model. As previously reported, SA-57 was considerably more potent in elevating anandamide (AEA) than 2-arachidonyl glycerol (2-AG) in brain. Its anti-allodynic effects required cannabinoid (CB)1 and CB2 receptors; however, only CB2 receptors were necessary for the anti-edematous effects in the carrageenan assay. Although high doses of SA-57 alone were required to produce antinociception, low doses of this compound, which elevated AEA and did not affect 2-AG brain levels, augmented the antinociceptive effects of morphine, but lacked cannabimimetic side effects. Because of the high abuse liability of opioids and implication of the endocannabinoid system in the reinforcing effects of opioids, the final experiment tested whether SA-57 would alter heroin seeking behavior. Strikingly, SA-57 reduced heroin-reinforced nose poke behavior and the progressive ratio break point for heroin. In conclusion, the results of the present study suggest that inhibition of endocannabinoid degradative enzymes represents a promising therapeutic approach to decrease effective doses of opioids needed for clinical pain control, and may also possess therapeutic potential to reduce opioid abuse.

Effects of repeated social defeat on adolescent mice on cocaine-induced CPP and self-administration in adulthood: integrity of the blood-brain barrier.

Social stress in adulthood enhances cocaine self-administration, an effect that has been related with an increase in extracellular signal-regulated kinase and p38α mitogen-activated protein kinase phosphorylation. A detrimental effect of cocaine on blood-brain barrier (BBB) integrity has also been reported. This study evaluates the effects of repeated social defeat (RSD) during adolescence on the reinforcing and motivational effects of cocaine in adult mice and the changes induced by RSD on BBB permeability. Cocaine self-administration, conditioned place preference and quantitative analysis of claudin-5, laminin, collagen-IV and IgG immunoreactivity took place 3 weeks after RSD. Mice socially defeated during adolescence developed conditioned place preference and exhibited reinstated preference with a non-effective dose of cocaine (1 mg/kg). RSD mice needed significantly more sessions than control animals for the preference induced by 25 mg/kg of cocaine to be extinguished. However, acquisition of cocaine self-administration (0.5 mg/kg per injection) was delayed in the RSD group. Mice exposed to RSD displayed significant changes in BBB structure in adulthood, with a marked reduction in expression of the tight junction protein claudin-5 and an increase in basal laminin degradation (reflected by a decrease in laminin and collagen-IV expression) in the nucleus accumbens and hippocampus. The detrimental effect induced by cocaine (25 mg/kg) on collagen-IV expression in the hippocampus was more pronounced in RSD mice. In summary, our findings suggest that stress and cocaine can increase the long-term vulnerability of the brain to subsequent environmental insults as a consequence of a sustained disruption of the BBB.

Finger chondroid syringoma as a hypoechoic subcutaneous nodule in ultrasound.

Chondroid syringoma (CS) is an uncommon, benign epithelial skin mixed tumor. It is often located in the head and neck and is unusual in other parts of the body. It may be seen as a skin or soft tissue tumor. We present findings on high-resolution ultrasound and histology in a case of benign CS located on the right index finger. High-resolution ultrasound showed a solid hypoechoic, well-defined subcutaneous mass, adjacent to the tendon. Complete surgical excision was performed, and histopathology demonstrated an apocrine mixed tumor (CS). Although CS histological findings are well described, radiological features have been reported only in few cases and mainly in magnetic resonance. Chondroid syringoma should be suspected by high-resolution ultrasound as a differential diagnosis for a solid slow-growing soft tissue nodule in a finger, especially if the lesion has no contact with the underlying tendon.

Overexpression of the CHRNA5/A3/B4 genomic cluster in mice increases the sensitivity to nicotine and modifies its reinforcing effects.

Nicotinic acetylcholine receptors (nAChRs) are ligand-gated pentameric ion channels that account for the effects of nicotine. Recent genetic studies have highlighted the importance of variants of the CHRNA5/A3/B4 genomic cluster in human nicotine dependence. Among these genetic variants those found in non-coding segments of the cluster may contribute to the pathophysiology of tobacco use through alterations in the expression of these genes. To discern the in vivo effects of the cluster, we generated a transgenic mouse overexpressing the human CHRNA5/A3/B4 cluster using a bacterial artificial chromosome. Transgenic mice showed increased functional α3ß4-nAChRs in brain regions where these subunits are highly expressed under normal physiological conditions. Moreover, they exhibited increased sensitivity to the pharmacological effects of nicotine along with higher activation of the medial habenula and reduced activation of dopaminergic neurons in the ventral tegmental area after acute nicotine administration. Importantly, transgenic mice showed increased acquisition of nicotine self-administration (0.015 mg/kg per infusion) and a differential response in the progressive ratio test. Our study provides the first in vivo evidence of the involvement of the CHRNA5/A3/B4 genomic cluster in nicotine addiction through modifying the activity of brain regions responsible for the balance between the rewarding and the aversive properties of this drug.

Characterization of the enzyme aac(3)-id in a clinical isolate of Salmonella enterica serovar Haifa causing traveler's diarrhea

Introducciónel objetivo de este estudio fue identificar el mecanismo de sensibilidad disminuida a gentamicina en un aislamiento clínico de Salmonella, lo que nos condujo a la detección de un gen que codifica una acetiltransferasa modificante de aminoglucósidos localizada en un integron tipo 1.Métodosla cepa multiresistente de Salmonella fue aislada de las heces de un viajero a Egipto. La susceptibilidad a 12 agentes antimicrobianos se determinó mediante Kirby-Bauer. La presencia de integron clase 1 se realizó mediante PCR. El producto de PCR amplificado del integrón fue recuperado y secuenciado para conocer los genes que contenía dicho integrón. Además se determinó la susceptibilidad a gentamicina C1a, gentamicina C1, sisomicina, neomicina, dibekacina, kanamicina, tobramicina, amikacina, netilmicina, apramicina, dactimicina, espectinomicina, estreptomicina, lividomicina y butirosina. El kit de expresión Champion™ pET101 Directional TOPO® fue utilizado para clonar y expresar el gen aac(3)-I.Resultadosel aislamiento fue identificado como Salmonella enterica serovariedad Haifa, el cual presentaba resistencia al ácido nalidixico, tetraciclina y sensibilidad disminuida a gentamicina. Se observó la presencia de un integron tipo 1 con un tamaño de 1,500bp en el que se encontraron dos genes (aac(3)-Id y aadA7). El análisis de la sensibilidad a diferentes aminoglucósidos de la cepa de E. coli TOP10F’ transformada con el vector que contenia el gen aac(3)-Id demostró resistencia a gentamicina C1a, gentamicina C1, y dactimicina, en concordancia con la presencia del enzima pero era susceptible a sisomicina. La secuencia de aminoácidos presentaba un 100% de identidad con el enzima AAC(3)-Id.Conclusiónla presencia del enzima AAC(3)-Id ha sido descrita por primera vez en S. Haifa(AU)

IntroductionThe objective of this investigation was to identify the mechanism of decreased susceptibility to gentamicin in a Salmonella clinical isolate, leading to the detection of a aminoglycoside acetyltransferase gene found in a class 1 integron.MethodsA multidrug-resistant Salmonella strain was recovered from feces of a traveler to Egypt. The antimicrobial susceptibility test to 12 antimicrobial agents was performed with the Kirby-Bauer method. The presence of class 1 integron was determined by PCR. The amplified product was recovered and sequenced in order to establish the genes carried. In addition, susceptibility to gentamicin C1a, gentamicin C1, sisomicin, neomycin, dibekacin, kanamycin, tobramycin, amikacin, netilmicin, apramycin, dactimicin, spectinomycin, streptomycin, lividomycin and butirosin, was established. The Champion™ pET101 Directional TOPO® Expression Kit was used to clone and express the aac(3)-I gene.ResultsThe isolate was identified as Salmonella enterica serovar Haifa, showing resistance to nalidixic acid, tetracycline and decreased susceptibility to gentamicin. One integron with a size circa 1,500bp, encoding an aac(3)-Id plus aadA7 genes was observed. The analysis of the susceptibility to different aminoglycosides in the E. coli TOP10F’ transformed with the vector carrying aac(3)-Id gene showed resistance to gentamicin C1a, gentamicin C1, and dactimicin, in accordance with the presence of this enzyme but, was susceptible to sisomicin. The homology of the amino acid and nucleotide sequences with the AAC(3)-Id enzyme was of 100%.ConclusionThe presence of the AAC(3)-Id enzyme was described for the first time in a S. Haifa(UA)